A new device for high-pressure freezing of cultured cell monolayer using 10-microm-thin stainless discs as both culture plate and specimen carrier.
High-pressure freezing (HPF) has been generally accepted as the most reliable method for cryofixation of biological samples, yielding a deep vitreous freezing. In recent cell biology, mammalian cultured cells are widely used, but HPF of cultured cell monolayer has not reached its full potential. In this study, we developed a new reliable device for HPF of cultured cell monolayer by using a 10-microm-thin stainless disc both as culture plate and specimen carrier. We describe the practical procedure, and demonstrate fine structures of HeLa cells cultured and cryofixed on the stainless discs as results.
Sawaguchi
Akira
A
Department of Anatomy, Ultrastructural Cell Biology, Faculty of Medicine, University of Miyazaki, Japan. sawa@fc.miyazaki-u.ac.jp
Aoyama
Fumiyo
F
Ide
Soyuki
S
Goto
Yoshiteru
Y
Suganuma
Tatsuo
T
D002478:Cells, Cultured / Q000648:ultrastructure*; D015925:Cryopreservation / Q000295:instrumentation / Q000379:methods; D004867:Equipment Design; D017110:Freeze Substitution; D006367:HeLa Cells; D006801:Humans; D008854:Microscopy, Electron; D011312:Pressure